GSM7711682: 17B_AAVS1_2stim; Homo sapiens; RNA-Seq
In both J-Lat and primary CD4+ T cell RNA isolation experiments, 0.1-2E6 cells were isolated and resuspended in 350 uL of RLT Plus (Qiagen, 1053393) + 1% 2-mercaptoethanol (Millipore Sigma, M3148). Cells were frozen in buffer RLT plus until ready to continue with isolation. Thawed RLT lysates were then run over a QIAshredder column (Qiagen, 79654) and subsequently over a gDNA eliminator column. Qiagen RNeasy Plus Mini Kit was then used in order to obtain purified total RNA. RNA was submitted for TapeStation RNA assay or HighSense RNA assay (Fred Hutch Core Facilities) and RINe scores were all found to be greater than or equal to 9.6. For J-Lat cells, library was prepared using Illumina TruSeq stranded mRNA standard protocol. For primary CD4+ T cells, library was prepared using Takara/Clontech SMARTv4 + Ilumina Nextera XT protocol.